EP3971202A1 - Film de résine formé d'un matériau d'échafaudage pour culture cellulaire, support de culture cellulaire et récipient de culture cellulaire - Google Patents

Film de résine formé d'un matériau d'échafaudage pour culture cellulaire, support de culture cellulaire et récipient de culture cellulaire Download PDF

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Publication number
EP3971202A1
EP3971202A1 EP20805342.1A EP20805342A EP3971202A1 EP 3971202 A1 EP3971202 A1 EP 3971202A1 EP 20805342 A EP20805342 A EP 20805342A EP 3971202 A1 EP3971202 A1 EP 3971202A1
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Prior art keywords
phase
resin film
cell culture
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resin
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German (de)
English (en)
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EP3971202A4 (fr
Inventor
Hiroki Iguchi
Yuuhei ARAI
Nobuhiko Inui
Mayumi YUKAWA
Daigo Kobayashi
Yuuta Nakamura
Kenta TAKAKURA
Satoshi Haneda
Ryoma ISHII
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Sekisui Chemical Co Ltd
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Sekisui Chemical Co Ltd
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Publication of EP3971202A1 publication Critical patent/EP3971202A1/fr
Publication of EP3971202A4 publication Critical patent/EP3971202A4/fr
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    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F20/00—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride, ester, amide, imide or nitrile thereof
    • C08F20/02—Monocarboxylic acids having less than ten carbon atoms, Derivatives thereof
    • C08F20/10—Esters
    • C08F20/12—Esters of monohydric alcohols or phenols
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/64—Cyclic peptides containing only normal peptide links
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F16/00—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal or ketal radical
    • C08F16/02—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal or ketal radical by an alcohol radical
    • C08F16/04—Acyclic compounds
    • C08F16/06—Polyvinyl alcohol ; Vinyl alcohol
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F261/00—Macromolecular compounds obtained by polymerising monomers on to polymers of oxygen-containing monomers as defined in group C08F16/00
    • C08F261/02—Macromolecular compounds obtained by polymerising monomers on to polymers of oxygen-containing monomers as defined in group C08F16/00 on to polymers of unsaturated alcohols
    • C08F261/04—Macromolecular compounds obtained by polymerising monomers on to polymers of oxygen-containing monomers as defined in group C08F16/00 on to polymers of unsaturated alcohols on to polymers of vinyl alcohol
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F261/00—Macromolecular compounds obtained by polymerising monomers on to polymers of oxygen-containing monomers as defined in group C08F16/00
    • C08F261/12—Macromolecular compounds obtained by polymerising monomers on to polymers of oxygen-containing monomers as defined in group C08F16/00 on to polymers of unsaturated acetals or ketals
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F8/00—Chemical modification by after-treatment
    • C08F8/30—Introducing nitrogen atoms or nitrogen-containing groups
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
    • C08J7/12—Chemical modification
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D151/00—Coating compositions based on graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Coating compositions based on derivatives of such polymers
    • C09D151/003—Coating compositions based on graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Coating compositions based on derivatives of such polymers grafted on to macromolecular compounds obtained by reactions only involving unsaturated carbon-to-carbon bonds
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/20—Material Coatings
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    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
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    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
    • C12M25/04—Membranes; Filters in combination with well or multiwell plates, i.e. culture inserts
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/14—Scaffolds; Matrices
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0068—General culture methods using substrates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1019—Tetrapeptides with the first amino acid being basic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F2810/00—Chemical modification of a polymer
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2351/00—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers
    • C08J2351/06—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers grafted on to homopolymers or copolymers of aliphatic hydrocarbons containing only one carbon-to-carbon double bond
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    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/20—Small organic molecules
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/30—Synthetic polymers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/30—Synthetic polymers
    • C12N2533/40—Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
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    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/50—Proteins

Definitions

  • the present invention relates to a resin film formed of a cell culture scaffold material.
  • the present invention also relates to a cell culture carrier and a cell culture vessel including the resin film.
  • hPSC human pluripotent stem cells
  • hESC human embryonic stem cells
  • hiPSC human induced pluripotent stem cells
  • Patent Document 1 discloses a cell culture carrier composed of a molded product made of a polyvinyl acetal compound or a molded product made of the polyvinyl acetal compound and a water-soluble polysaccharide, in which the polyvinyl acetal compound has a degree of acetalization of 20 to 60 mol%.
  • Patent Document 2 discloses a composition containing a first fiber polymer scaffolding, in which the fibers of the first fiber polymer scaffolding are aligned. It is described that the fibers constituting the first fiber polymer scaffolding are composed of an aliphatic polyester such as polyglycolic acid or polylactic acid.
  • Patent Document 3 describes a cell culture method for maintaining undifferentiation of pluripotent stem cells, including a step of culturing the pluripotent stem cells on an incubator having a surface coated with a polyrotaxan block copolymer.
  • a natural polymer material when used as a scaffold material, fixation of cells after seeding can be enhanced.
  • fixation of cells after seeding is extremely high when an adhesive protein such as laminin or vitronectin, or a matrigel derived from mouse sarcoma is used as a natural polymer.
  • an adhesive protein such as laminin or vitronectin, or a matrigel derived from mouse sarcoma is used as a natural polymer.
  • natural polymer materials are expensive, have large variations between lots because they are naturally derived substances, or have safety concerns due to animal-derived components.
  • the scaffold materials using synthetic resins have good operability, are inexpensive, have less variation between lots and are excellent in safety, in comparison to scaffold materials using natural polymer materials.
  • the synthetic resins swell excessively because synthetic resins having high hydrophilicity are used.
  • synthetic resins have lower affinity for cells than natural polymer materials, a cell mass may exfoliate during culturing.
  • the scaffolding materials using synthetic resins have low fixation of cells after seeding, and the cells may not proliferate sufficiently.
  • An object of the present invention is to provide a resin film formed of a cell culture scaffold material, a cell culture carrier, and a cell culture vessel, which have excellent fixation of cells after seeding and are capable of enhancing proliferation rate of cells.
  • the cell culture scaffold material contains a synthetic resin
  • the resin film has a phase-separated structure including at least a first phase and a second phase, and a ratio of the surface area of one of the first phase and the second phase to the entire surface is 0.01 or more and 0.95 or less.
  • the ratio of the peripheral length to the area of the second phase is 0.001 (1/nm) or more and 0.40 (1/nm) or less.
  • the phase-separated structure is a sea-island structure, the first phase is a sea part, and the second phase is an island part.
  • the number of the second phase as an island part is 1 piece/ ⁇ m 2 or more and 5,000 pieces/ ⁇ m 2 or less.
  • the phase-separated structure is composed of a phase-separated structure within a molecule of the synthetic resin.
  • a dispersion term component of surface free energy is 25.0 mJ/m 2 or more and 50.0 mJ/m 2 or less
  • a polar term component of surface free energy is 1.0 mJ/m 2 or more and 20.0 mJ/m 2 or less.
  • the synthetic resin has a cationic functional group, and the content of the cationic functional group contained in a structural unit of the synthetic resin is 0.2 mol% or more and 50 mol% or less.
  • the second phase has a peptide portion.
  • the peptide portion has a cell-adhesive amino acid sequence.
  • the resin film has a water swelling ratio of 50% or less.
  • the resin film has a storage elastic modulus at 100°C of 1.0 ⁇ 10 4 Pa or more and 1.0 ⁇ 10 8 Pa or less, and a ratio of a storage elastic modulus at 25°C to a storage elastic modulus at 100°C ((storage elastic modulus at 25°C)/(storage elastic modulus at 100°C)) of 1.0 ⁇ 10 1 or more and 1.0 ⁇ 10 5 or less.
  • the cell culture scaffold material does not substantially contain animal-derived raw materials.
  • the synthetic resin contains a vinyl polymer.
  • the synthetic resin contains at least a polyvinyl alcohol derivative or a poly(meth)acrylic acid ester.
  • the cell culture carrier according to the present invention includes a carrier, and a resin film constituted according to the present invention, and the resin film is arranged on a surface of the carrier.
  • the cell culture vessel according to the present invention includes a vessel body, and a resin film constituted according to the present invention, and the resin film is arranged on a surface of the vessel body.
  • a resin film formed of a cell culture scaffold material, a cell culture carrier, and a cell culture vessel which have excellent fixation of cells after seeding and are capable of enhancing proliferation rate of cells.
  • the present invention relates to a resin film formed of a cell culture scaffold material.
  • the cell culture scaffold material contains a synthetic resin.
  • the resin film of the present invention has a phase-separated structure including at least a first phase and a second phase.
  • a ratio of the surface area of one of the first phase and the second phase to the entire surface is 0.01 or more and 0.95 or less.
  • the resin film of the present invention has the above structure, it has excellent fixation of cells after seeding and is capable of enhancing proliferation rate of cells.
  • Conventional cell culture scaffold materials using natural polymer materials can enhance fixation of cells after seeding, but are expensive, have large variations between lots because they are naturally derived substances, or have safety concerns due to animal-derived components.
  • the synthetic resins swell excessively or have low affinity for cells, so that a cell mass may exfoliate during culturing. Therefore, the conventional scaffolding materials using synthetic resins have low fixation of cells after seeding, and the cells may not proliferate sufficiently.
  • the present inventors have focused on a phase-separated structure of a resin film formed of a cell culture scaffold material, and have found that a phase-separated structure having a ratio of the surface area of one of the first phase and the second phase to the entire surface in the above specific range can enhance the affinity for cells, thereby capable of enhancing adhesion after seeding, and thus enhancing the proliferation rate of cells.
  • a reason for this is not clear, but when having such a phase-separated structure, energy distribution proceeds smoothly, and positions and ratios of the first phase and the second phase having different affinities and intensities can be adjusted. Therefore, it is considered that the affinity can be enhanced regardless of the type of cell, and accumulation and adsorption effect of cells or cell surface proteins can be realized.
  • the resin film of the present invention adhesiveness to the cells after seeding can be enhanced, and the proliferation rate of cells can be enhanced.
  • the synthetic resin since the synthetic resin can be used as described above, it has good operability, is inexpensive, has less variation between lots and is excellent in safety, in comparison to scaffold materials using natural polymer materials.
  • a synthetic resin having a peptide portion may be used as the synthetic resin. Details of the synthetic resin having a peptide portion will be described later.
  • the ratio of the surface area of one of the first phase and the second phase to the entire surface is 0.01 or more, preferably 0.10 or more, 0.95 or less, and more preferably 0.90 or less.
  • the surface area fraction is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • examples of the phase-separated structure include microphase-separated structures such as a sea-island structure, a cylinder structure, a gyroid structure, or a lamellar structure.
  • the first phase can be a sea part and the second phase can be an island part.
  • the cylinder structure, gyroid structure, or lamellar structure for example, a phase having a largest surface area can be the first phase, and a phase having a second largest surface area can be the second phase.
  • the sea-island structure is preferable as the phase-separated structure.
  • by having a continuous phase and a discontinuous phase it is possible to enhance the affinity for cells and further enhance the adhesiveness to the cells after seeding, thereby further enhancing the proliferation rate of cells.
  • the surface area fraction of the second phase to the entire surface is 0.01 or more, preferably 0.1 or more, and more preferably 0.2 or more, and 0.95 or less, preferably 0.9 or less, and more preferably 0.8 or less.
  • the surface area fraction is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • a ratio of the peripheral length to the area of the second phase is preferably 0.001 (1/nm) or more, more preferably 0.0015 (1/nm) or more, and further preferably 0.008 (1/nm) or more.
  • the ratio of the peripheral length to the area of the second phase (peripheral length/area) is preferably 0.40 (1/nm) or less, more preferably 0.20 (1/nm) or less, further preferably 0.08 (1/nm) or less, and particularly preferably 0.013 (1/nm) or less.
  • the ratio of the peripheral length to the area of the second phase is preferably 0.001 (1/nm) or more, more preferably 0.0015 (1/nm) or more, preferably 0.08 (1/nm) or less, and more preferably 0.013 (1/nm) or less.
  • the ratio (peripheral length/area) is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the ratio of the peripheral length to the area of the second phase is preferably 0.008 (1/nm) or more, more preferably 0.013 (1/nm) or more, preferably 0.40 (1/nm) or less, more preferably 0.20 (1/nm) or less, and further preferably 0.10 (1/nm) or less.
  • the ratio (peripheral length/area) is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the number of the second phases as island parts is preferably 1 piece/ ⁇ m 2 or more, more preferably 2 pieces/ ⁇ m 2 or more, further preferably 10 pieces/ ⁇ m 2 or more, preferably 5,000 pieces/ ⁇ m 2 or less, more preferably 1,000 pieces/ ⁇ m 2 or less, further preferably 500 pieces/ ⁇ m 2 or less, and particularly preferably 300 pieces/ ⁇ m 2 or less.
  • the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the average diameter of the second phases as island parts is preferably 20 nm or more, more preferably 30 nm or more, further preferably 50 nm or more, particularly preferably 80 nm or more, preferably 3.5 ⁇ m or less, more preferably 3.0 ⁇ m or less, and further preferably 1.5 ⁇ m or less.
  • the average diameter of the second phases is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the average diameter of the second phases as island parts is preferably 50 nm or more, more preferably 100 nm or more, further preferably 120 nm or more, particularly preferably 200 nm or more, preferably 1 ⁇ m or less, more preferably 300 nm or less, and further preferably 250 nm or less.
  • the average diameter of the second phases is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the average diameter of the second phases as island parts is preferably 10 nm or more, more preferably 20 nm or more, further preferably 40 nm or more, preferably 1 ⁇ m or less, more preferably 300 nm, and further preferably 100 nm or less.
  • the average diameter of the second phases is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • phase-separated structure The presence or absence of a phase-separated structure and parameters indicating the phase-separated structure as described above can be confirmed by, for example, an atomic force microscope (AFM), a transmission electron microscope (TEM), a scanning electron microscope (SEM), or the like.
  • AFM atomic force microscope
  • TEM transmission electron microscope
  • SEM scanning electron microscope
  • the ratio of the surface area of one of the first phase and the second phase to the entire surface can be obtained from the above-mentioned microscopic observation image using image analysis software such as ImageJ.
  • the ratio of the surface area of one of the first phase and the second phase to the entire surface is obtained by, within an observation region (30 ⁇ m ⁇ 30 ⁇ m), dividing a surface area occupied by one of the first phase and the second phase by an area of the observation region.
  • the ratio of the peripheral length to the area of the second phase is obtained by, within the observation region (30 ⁇ m ⁇ 30 ⁇ m), dividing a total peripheral length of the second phase by a total area of the second phase.
  • the number of second phases as island parts can be obtained by dividing the number of second phases in the observation region (30 ⁇ m ⁇ 30 ⁇ m) by the area of the observation region. Also, the average diameter of the second phases as island parts is obtained as an average diameter of circles of the same area.
  • phase-separated structure as described above can be obtained by, for example, blending at least two different types of polymers, copolymerizing, graft-copolymerizing, or using a synthetic resin having a peptide portion to form a phase-separated structure between molecules or within a molecule of the synthetic resin.
  • the phase-separated structure is formed of a phase-separated structure within a molecule.
  • the synthetic resin is preferably a copolymer of at least two different types of polymers or a synthetic resin having a peptide portion, and is more preferably a graft copolymer or a synthetic resin having a peptide portion.
  • phase-separated structure as described above is preferably obtained by copolymerizing two or more different types of polymers (monomers) having a solubility parameter (SP value) of 0.1 or more, preferably 0.5 or more, and more preferably 1 or more.
  • SP value solubility parameter
  • the sea-island structure can be formed more easily.
  • the SP value is a measure of intermolecular force acting between a solvent and a solute, and is a measure of affinity between substances.
  • a unit of SP value is (cal/cm 3 ) 0.5 .
  • the Fedors method is described in Journal of the Adhesion Society of Japan, 1986, Vol. 22, p. 566 .
  • phase separation parameters indicating the phase-separated structure such as the surface area fraction can be adjusted by, for example, controlling a blending ratio of the two types of polymers or structure of the polymers, or controlling the content of the peptide portion.
  • phase different from the first phase and the second phase there may be other phase different from the first phase and the second phase.
  • the other phase may be one phase or a plurality of phases.
  • Such a phase can be obtained, for example, by copolymerization such as grafting still another polymer (monomer) having a different SP value.
  • two phases occupying large areas of the surface of the resin film are defined as the first phase and the second phase.
  • a dispersion term component of surface free energy in the resin film formed of the cell culture scaffold material is preferably 25.0 mJ/m 2 or more and 50.0 mJ/m 2 or less.
  • hydrophilicity of the cell culture scaffold material can be appropriately adjusted, and by a synergistic effect with the phase-separated structure, interfacial adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the dispersion term component is more preferably 30.0 mJ/m 2 or more, further preferably 35.0 mJ/m 2 or more, more preferably 47.0 mJ/m 2 or less, and further preferably 45.5 mJ/m 2 or less.
  • a polar term component of surface free energy in the resin film formed of the cell culture scaffold material is preferably 1.0 mJ/m 2 or more and 20.0 mJ/m 2 or less.
  • the polar term component is more preferably 2.0 mJ/m 2 or more, further preferably 3.0 mJ/m 2 or more, more preferably 10.0 mJ/m 2 or less, and further preferably 5.0 mJ/m 2 or less.
  • Dispersion term component ⁇ d of surface free energy and dipole component ⁇ p as a polar term component of surface free energy are calculated using the Kaelble-Uy theoretical formula.
  • the Kaelble-Uy theoretical formula is a theoretical formula based on an assumption total surface free energy ⁇ is a sum of the dispersion term component ⁇ d and the dipole component ⁇ p , as shown by Formula (1) below.
  • contact angle ⁇ with respect to the resin film formed of the cell culture scaffold material is measured using two types of liquids whose surface free energy of liquid ⁇ l is known, and simultaneous equations of ⁇ s d and ⁇ s p are solved, whereby the dispersion term component ⁇ d and dipole component ⁇ p of surface free energy of the resin film formed of the cell culture scaffold material can be obtained.
  • the contact angle ⁇ is measured as follows using a contact angle meter (for example, "DMo-701" manufactured by Kyowa Interface Science Co., Ltd.).
  • Pure water or diiodomethane (1 ⁇ L) is added dropwise to a surface of the resin film formed of the cell culture scaffold material.
  • An angle formed by the pure water 30 seconds after dropping and the resin film is defined as contact angle ⁇ with respect to the pure water.
  • an angle formed by the diiodomethane 30 seconds after dropping and the resin film is defined as contact angle ⁇ with respect to the diiodomethane.
  • the dispersion term component ⁇ d of the surface free energy can be reduced.
  • the dipole component ⁇ p of the surface free energy can be reduced.
  • a storage elastic modulus at 100°C is preferably 0.6 ⁇ 10 4 Pa or more, more preferably 0.8 ⁇ 10 4 Pa or more, further preferably 1.0 ⁇ 10 4 Pa or more, preferably 1.0 ⁇ 10 8 Pa or less, more preferably 0.8 ⁇ 10 8 Pa or less, and further preferably 1.0 ⁇ 10 7 Pa or less.
  • the resin film formed from the cell culture scaffold material of the present invention has a ratio of a storage elastic modulus at 25°C to a storage elastic modulus at 100°C ((storage elastic modulus at 25°C)/(storage elastic modulus at 100°C)) is preferably 1.0 ⁇ 10 1 or more, more preferably 5.0 ⁇ 10 1 or more, further preferably 8.0 ⁇ 10 2 or more, preferably 1.0 ⁇ 10 5 or less, more preferably 0.75 ⁇ 10 5 or less, and further preferably 0.5 ⁇ 10 5 or less.
  • fixation of cells after seeding can be further enhanced.
  • the storage elastic moduli at 25°C and 100°C are measured, for example, by a dynamic viscoelasticity measuring device (manufactured by IT Keisoku Seigyo Co., Ltd., DVA-200), under tensile conditions at a frequency of 10 Hz and a temperature range of -150°C to 150°C at a heating rate of 5°C/min.
  • the storage elastic moduli at 25°C and 100°C are obtained from a graph of the obtained tensile storage elastic modulus, and 25°C storage elastic modulus/100°C storage elastic modulus is calculated.
  • the measurement is performed using a measurement sample with a length of 50 mm, a width of 5 to 20 mm, and a thickness of 0.1 to 1.0 mm, under conditions of 10 Hz, a strain of 0.1%, a temperature of -150°C to 150°C, and a heating rate of 5°C/min.
  • the storage elastic moduli at 25°C and 100°C can be increased, for example, by increasing the degree of crosslinking in the synthetic resin, stretching the synthetic resin, and the like. Further, the storage elastic moduli at 25°C and 100°C can be lowered by reducing the number average molecular weight of the synthetic resin, lowering the glass transition temperature, and the like.
  • the resin film formed of the cell culture scaffold material of the present invention has a water swelling ratio of preferably 50% or less and more preferably 40% or less. In this case, the fixation of cells after seeding can be further enhanced.
  • the lower limit of the water swelling ratio is not particularly limited, but can be set to, for example, 0.5%.
  • the water swelling ratio can be reduced, for example, by increasing the hydrophobic functional groups of the synthetic resin, reducing the number average molecular weight, and the like.
  • the cell culture scaffold material contains a synthetic resin (hereinafter, may be referred to as synthetic resin X).
  • a main chain of the synthetic resin X is preferably a carbon chain.
  • a "structural unit” refers to a repeating unit of a monomer constituting the synthetic resin.
  • the synthetic resin has a graft chain, it contains a repeating unit of a monomer constituting the graft chain.
  • the synthetic resin X When the synthetic resin X does not have a peptide portion, the synthetic resin X preferably has a cationic functional group.
  • the synthetic resin X having a peptide portion may or may not have a cationic functional group in a structural part other than the peptide portion.
  • the cationic functional group include substituents having a structure such as an amino group, an imino group, or an amide group.
  • Examples include, without particular limitation, conjugated amine-based functional groups such as hydroxyamino group, urea group, guanidine and biguanide, heterocyclic amino functional groups such as piperazine, piperidine, pyrrolidine, 1,4-diazabicyclo[2.2.2]octane, hexamethylenetetramine, morpholine, pyridine, pyridazine, pyrimidine, pyrazine, pyrrole, azatropylidene, pyridone, imidazole, benzimidazole, benzotriazole, pyrazole, oxazole, imidazoline, triazole, thiazole, thiazine, tetrazole, indole, isoindole, purine, quinoline, isoquinoline, quinazoline, quinoxaline, cinnoline, pteridine, carbazole, acridine, adenine,
  • the content of the cationic functional group contained in a structural unit of the synthetic resin X is preferably 0.2 mol% or more, preferably 2 mol% or more, more preferably 3 mol% or more, 50 mol% or less, preferably 10 mol% or less, and more preferably 7 mol% or less.
  • the synthetic resin X containing a cationic functional group within such a range, the fixation of cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the content of the cationic functional group can be measured by, for example, 1 H-NMR (nuclear magnetic resonance spectrum).
  • the synthetic resin X preferably contains a vinyl polymer, and more preferably is a vinyl polymer.
  • the vinyl polymer is a polymer of a compound having a vinyl group or a vinylidene group.
  • the synthetic resin X is a vinyl polymer, it is possible to more easily suppress swelling of the cell culture scaffold material in water.
  • the vinyl polymer include polyvinyl alcohol derivatives, poly(meth)acrylic acid esters, polyvinylpyrrolidone, polystyrene, ethylene-vinyl acetate copolymers, and the like.
  • the vinyl polymer is preferably a polyvinyl alcohol derivative or a poly(meth)acrylic acid ester, from the viewpoint of easily enhancing the adhesiveness to cells.
  • the cell culture scaffold material preferably contains synthetic resin X having a polyvinyl acetal skeleton.
  • the synthetic resin X having a polyvinyl acetal skeleton is preferably a copolymer of a structural unit of a polyvinyl acetal resin and a vinyl compound and/or a vinylidene compound.
  • the vinyl compound or vinylidene compound may be a vinyl polymer which is a polymer thereof.
  • the vinyl compound, vinylidene compound and vinyl polymer copolymerized with the polyvinyl acetal resin may be collectively referred to as "vinyl compound A".
  • the copolymer may be a block copolymer of a polyvinyl acetal resin and vinyl compound A, or may be a graft copolymer obtained by grafting vinyl compound A on a polyvinyl acetal resin.
  • the copolymer is preferably a graft copolymer. In this case, the phase-separated structure can be formed more easily.
  • vinyl compound and vinylidene compound examples include ethylene, allylamine, vinylpyrrolidone, maleic anhydride, maleimide, itaconic acid, (meth)acrylic acid, vinylamine, (meth)acrylic acid esters, and the like.
  • vinyl compounds only one type may be used, or two or more types may be used in combination. Therefore, it may be a vinyl polymer in which these vinyl compounds are copolymerized.
  • difference in SP value between the polyvinyl acetal resin and the vinyl compound A is preferably 0.5 or more. In this case, the phase-separated structure can be formed more easily.
  • the difference in SP value between the polyvinyl acetal resin and the vinyl compound A is more preferably 1.0 or more.
  • the upper limit of the difference in SP values is not particularly limited, but can be set to, for example, 10.0.
  • the first phase is a polyvinyl acetal resin and the second phase is vinyl compound A. It is preferable that the first phase is formed by a polyvinyl acetal resin part of the copolymer and the second phase is formed by a vinyl compound A part. In this case, it is preferable that the first phase of the polyvinyl acetal resin is a sea part and the second phase of the vinyl compound A is an island part. However, the first phase of the vinyl compound A may be a sea part, and the second phase of the polyvinyl acetal resin may be an island part.
  • the content fraction (mol/mol) of the vinyl compound A in the copolymer is preferably 0.015 or more, more preferably 0.3 or more, preferably 0.95 or less, more preferably 0.90 or less, and further preferably 0.70 or less.
  • the content fraction is the above lower limit or more, the phase-separated structure can be more easily formed.
  • the content fraction is the upper limit or less, the proliferation rate of cells can be further increased.
  • polyvinyl acetal resin polyvinyl acetal resin part of the copolymer
  • the polyvinyl acetal resin has an acetal group, an acetyl group, and a hydroxyl group in side chains.
  • a method for synthesizing a polyvinyl acetal resin includes at least a step of acetalizing polyvinyl alcohol with an aldehyde.
  • the aldehyde used for acetalizing polyvinyl alcohol to obtain a polyvinyl acetal resin is not particularly limited.
  • Examples of the aldehyde include aldehydes having 1 to 10 carbon atoms.
  • the aldehyde may have a chain aliphatic group, a cyclic aliphatic group or an aromatic group.
  • the aldehyde may be a chain aldehyde or a cyclic aldehyde.
  • aldehyde examples include formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, pentanal, hexanal, heptanal, octanal, nonanal, decanal, acrolein, benzaldehyde, cinnamaldehyde, perillaldehyde, formylpyridine, formylimidazole, formylpyrrole, formylpiperidine, formyltriazole, formyltetrazole, formylindole, formylisoindole, formylpurine, formylbenzimidazole, formylbenzotriazole, formylquinoline, formylisoquinoline, formylquinoxaline, formylcinnoline, formylpteridine, formylfuran, formyloxolane, formyloxane, formylthiophene, formylthio
  • the aldehyde is preferably formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, or pentanal, and more preferably butyraldehyde. Therefore, the polyvinyl acetal skeleton is preferably a polyvinyl butyral skeleton.
  • the polyvinyl acetal resin is preferably a polyvinyl butyral resin.
  • the polyvinyl acetal resin preferably has a Bronsted basic group or a Bronsted acidic group, and more preferably has a Bronsted basic group. That is, it is preferable that a part of the polyvinyl acetal resin is modified with a Bronsted basic group or a Bronsted acidic group, and more preferably a part of the polyvinyl acetal resin is modified with a Bronsted basic group.
  • the Bronsted basic group is a generic term for a functional group that can receive a hydrogen ion H + from another substance.
  • Examples of the Bronsted basic group include amine-based basic groups such as a substituent having an imine structure, a substituent having an imide structure, a substituent having an amine structure, or a substituent having an amide structure.
  • Bronsted basic group examples include, without particular limitation, conjugated amine-based functional groups such as hydroxyamino group, urea group, guanidine and biguanide, heterocyclic amino functional groups such as piperazine, piperidine, pyrrolidine, 1,4-diazabicyclo[2.2.2]octane, hexamethylenetetramine, morpholine, pyridine, pyridazine, pyrimidine, pyrazine, pyrrole, azatropylidene, pyridone, imidazole, benzimidazole, benzotriazole, pyrazole, oxazole, imidazoline, triazole, thiazole, thiazine, tetrazole, indole, isoindole, purine, quinoline, isoquinoline, quinazoline, quinoxaline, cinnoline, pteridine, carbazole, acridine,
  • Examples of the Bronsted acidic group include a carboxyl group, a sulfonic acid group, a maleic acid group, a sulfinic acid group, a sulfenic acid group, a phosphoric acid group, a phosphonic acid group, salts thereof, and the like.
  • the Bronsted acidic group is preferably a carboxyl group.
  • the polyvinyl acetal resin preferably has a structural unit having an imine structure, a structural unit having an imide structure, a structural unit having an amine structure, or a structural unit having an amide structure.
  • the polyvinyl acetal resin may have only one type of these structural units, or may have two or more types.
  • the polyvinyl acetal resin may have a structural unit having an imine structure.
  • the polyvinyl acetal resin preferably has an imine structure in a side chain.
  • the polyvinyl acetal resin may have a structural unit having an imide structure.
  • the polyvinyl acetal resin preferably has an imino group in a side chain.
  • the imino group may be directly bonded to a carbon atom constituting a main chain of the polyvinyl acetal resin, or may be bonded to the main chain via a linking group such as an alkylene group.
  • the polyvinyl acetal resin may have a structural unit having an amine structure.
  • the amine group in the amine structure may be a primary amine group, a secondary amine group, a tertiary amine group, or a quaternary amine group.
  • the structural unit having an amine structure may be a structural unit having an amide structure.
  • the polyvinyl acetal resin preferably has an amine structure or an amide structure in a side chain.
  • the amine structure or the amide structure may be directly bonded to the carbon atom constituting a main chain of the polyvinyl acetal resin, or may be bonded to the main chain via a linking group such as an alkylene group.
  • the content of the structural unit having an imine structure, the content of the structural unit having an imide structure, the content of the structural unit having an amine structure, and the content of the structural unit having an amide structure can be measured by 1 H-NMR (nuclear magnetic resonance spectrum).
  • the vinyl compound A is preferably a (meth)acrylic acid ester or a poly(meth)acrylic acid ester resin.
  • the synthetic resin X is a copolymer obtained by graft-copolymerizing a polyvinyl acetal resin with a (meth)acrylic acid ester or a poly(meth)acrylic acid ester resin which is a polymer thereof.
  • the poly(meth)acrylic acid ester resin can be obtained by polymerizing the (meth)acrylic acid ester or by polymerizing the (meth)acrylic acid ester with the other monomers.
  • Examples of the (meth)acrylic acid ester include (meth)acrylic acid alkyl ester, (meth)acrylic acid cyclic alkyl ester, (meth)acrylic acid aryl ester, (meth)acryl amides, polyethylene glycol (meth)acrylates, phosphorylcholine (meth)acrylate, and the like.
  • Examples of the (meth)acrylic acid alkyl ester include methyl (meth)acrylate, ethyl (meth)acrylate, n-propyl (meth)acrylate, isopropyl (meth)acrylate, n-butyl (meth)acrylate, isobutyl (meth)acrylate, t-butyl (meth)acrylate, n-octyl (meth)acrylate, isooctyl (meth)acrylate, 2-ethylhexyl (meth)acrylate, nonyl (meth)acrylate, isononyl (meth)acrylate, decyl (meth)acrylate, isodecyl (meth)acrylate, lauryl (meth)acrylate, stearyl (meth)acrylate, isotetradecyl (meth)acrylate, and the like.
  • the (meth)acrylic acid alkyl ester may be substituted with a substituent such as an alkoxy group having 1 to 3 carbon atoms and a tetrahydrofurfuryl group.
  • a substituent such as an alkoxy group having 1 to 3 carbon atoms and a tetrahydrofurfuryl group.
  • Examples of such (meth)acrylic acid alkyl ester include methoxyethyl acrylate, tetrahydrofurfuryl acrylate, and the like.
  • Examples of the (meth)acrylic acid cyclic alkyl ester include cyclohexyl (meth)acrylate, isobornyl (meth)acrylate, and the like.
  • Examples of the (meth)acrylic acid aryl ester include phenyl (meth)acrylate, benzyl (meth)acrylate, and the like.
  • Examples of the (meth)acrylamides include (meth)acrylamide, N-isopropyl (meth)acrylamide, N-tert-butyl (meth)acrylamide, N,N'-dimethyl (meth)acrylamide, (3-(meth)acrylamide propyl)trimethylammonium chloride, 4-(meth)acryloylmorpholine, 3-(meth)acryloyl-2-oxazolidinone, N-[3-(dimethylamino)propyl](meth)acrylamide, N-(2-hydroxyethyl)(meth)acrylamide, N-methylol(meth)acrylamide, 6-(meth)acrylamide hexane acid, and the like.
  • polyethylene glycol (meth)acrylates examples include methoxy-polyethylene glycol (meth)acrylate, ethoxy-polyethylene glycol (meth)acrylate, hydroxy-polyethylene glycol (meth)acrylate, methoxy-diethylene glycol (meth)acrylate, ethoxy-diethylene glycol (meth)acrylate, hydroxy-diethylene glycol (meth)acrylate, methoxytriethylene glycol (meth)acrylate, ethoxy-triethylene glycol (meth)acrylate, hydroxy-triethylene glycol (meth)acrylate, and the like.
  • Examples of the phosphorylcholine (meth)acrylate include 2-(meth)acryloyloxyethyl phosphorylcholine and the like.
  • a vinyl compound is preferably used as the other monomer copolymerized with the (meth)acrylic acid ester.
  • the vinyl compound include ethylene, allylamine, vinylpyrrolidone, vinylimidazole, maleic anhydride, maleimide, itaconic acid, (meth)acrylic acid, vinylamine, (meth)acrylic acid ester, and the like.
  • the vinyl compound only one type may be used, or two or more types may be used in combination.
  • (meth)acrylic means “acrylic” or “methacrylic
  • (meth)acrylate means “acrylate” or “methacrylate”.
  • the synthetic resin X may be a copolymer of a resin having a poly(meth)acrylic acid ester skeleton and other vinyl compound as long as the phase-separated structure of the present invention can be formed.
  • ethylene, allylamine, vinylpyrrolidone, maleic anhydride, maleimide, itaconic acid, (meth)acrylic acid, vinylamine, or other (meth)acrylic acid ester having a different SP value can be used.
  • the cell culture scaffold material preferably contains synthetic resin X having a peptide portion.
  • the synthetic resin X having a peptide portion can be obtained by reacting the synthetic resin X with a linker and a peptide.
  • the synthetic resin X having a peptide portion is preferably a peptide-conjugated polyvinyl acetal resin having a polyvinyl acetal resin portion, a linker portion, and a peptide portion, and more preferably a peptide-conjugated polyvinyl butyral resin having a polyvinyl butyral resin portion, a linker portion, and a peptide portion.
  • As the synthetic resin X having a peptide portion only one type may be used, or two or more types may be used in combination.
  • the peptide portion is preferably composed of 3 or more amino acids, more preferably composed of 4 or more amino acids and further preferably composed of 5 or more amino acids, and is preferably composed of 10 or less amino acids and more preferably composed of 6 or less amino acids.
  • the number of amino acids constituting the peptide portion is the above lower limit or more and the above upper limit or less, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • the peptide portion preferably has a cell-adhesive amino acid sequence.
  • the cell-adhesive amino acid sequence refers to an amino acid sequence whose cell adhesion activity has been confirmed by phage display method, sepharose beads method, or plate coating method.
  • phage display method for example, a method described in " The Journal of Cell Biology, Volume 130, Number 5, September 1995 1189-1196 " can be used.
  • sepharose beads method for example, a method described in " Protein, Nucleic Acid and Enzyme, Vol.45 No.15 (2000) 2477 " can be used.
  • As the plate coating method for example, a method described in " Protein, Nucleic Acid and Enzyme, Vol.45 No.15 (2000) 2477 " can be used.
  • cell-adhesive amino acid sequence examples include RGD sequence (Arg-Gly-Asp), YIGSR sequence (Tyr-Ile-Gly-Ser-Arg), PDSGR sequence (Pro-Asp-Ser-Gly-Arg), HAV sequence (His-Ala-Val), ADT sequence (Ala-Asp-Thr), QAV sequence (Gln-Ala-Val), LDV sequence (Leu-Asp-Val), IDS sequence (Ile-Asp-Ser), REDV sequence (Arg-Glu-Asp-Val), IDAPS sequence (Ile-Asp-Ala-Pro-Ser), KQAGDV sequence (Lys-Gln-Ala-Gly-Asp-Val), TDE sequence (Thr-Asp-Glu), and the like.
  • examples of the cell-adhesive amino acid sequence include sequences described in “ Medicina Philosophica, Vol. 9, No. 7, pp. 527-535, 1990 " and “ Journal of Osaka Women's and Children's Hospital, Vol. 8, No. 1, pp. 58-66, 1992 ", and the like.
  • the peptide portion may have only one type of cell-adhesive amino acid sequence, or may have two or more types.
  • the cell-adhesive amino acid sequence preferably has at least one of the above-mentioned cell-adhesive amino acid sequences, more preferably has at least an RGD sequence, a YIGSR sequence or a PDSGR sequence, and further preferably has at least an RGD sequence represented by the following formula (1).
  • the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of cells can be further enhanced.
  • X represents Gly, Ala, Val, Ser, Thr, Phe, Met, Pro, or Asn.
  • the peptide portion may be linear or may have a cyclic peptide skeleton.
  • the cyclic peptide skeleton is a cyclic skeleton composed of a plurality of amino acids. From the viewpoint of more effectively exhibiting the effect of the present invention, the cyclic peptide skeleton is preferably composed of 4 or more amino acids, preferably composed of 5 or more amino acids, and preferably composed of 10 or less amino acids.
  • the content of the peptide portion is preferably 0.1 mol% or more, more preferably 1 mol% or more, further preferably 5 mol% or more, and particularly preferably 10 mol% or more.
  • the content of the peptide portion is preferably 60 mol% or less, more preferably 50 mol% or less, further preferably 35 mol% or less, and particularly preferably 25 mol% or less.
  • the content (mol%) of the peptide portion is the amount of substance of the peptide portion with respect to the total of the amount of substance of structural units constituting the synthetic resin X having a peptide portion.
  • the content of the peptide portion can be measured by FT-IR or LC-MS.
  • the second phase has a peptide portion, and it is more preferable that the peptide portion has the cell-adhesive amino acid sequence. It is preferable that the second phase is formed by the peptide portion of the synthetic resin X having a peptide portion. In this case, it is preferable that the second phase having a peptide portion is an island part. However, the first phase may have a peptide portion, and the second phase having a peptide portion may be a sea part.
  • the synthetic resin X having a peptide portion it is preferable that the synthetic resin X part and the peptide portion are bonded via a linker. That is, the synthetic resin X having a peptide portion is preferably a synthetic resin X having a peptide portion and a linker portion.
  • the linker only one type may be used, or two or more types may be used in combination.
  • the linker is preferably a compound having a functional group capable of condensing with the carboxyl group or amino group of the peptide.
  • the functional group capable of condensing with the carboxyl group or amino group of the peptide include a carboxyl group, a thiol group, an amino group, and the like.
  • the linker is preferably a compound having a carboxyl group.
  • the above-mentioned vinyl compound A can also be used.
  • Examples of the linker having a carboxyl group include (meth)acrylic acid, a carboxyl group-containing acrylamide, and the like.
  • a carboxylic acid having a polymerizable unsaturated group (carboxylic acid monomer)
  • the carboxylic acid monomer can be polymerized by graft polymerization when the linker and the synthetic resin X are reacted, so that the number of the carboxyl groups capable of reacting with a peptide can be increased.
  • the cell culture scaffold material contains the synthetic resin X.
  • the content of the synthetic resin X in 100% by weight of the cell culture scaffold material is preferably 90% by weight or more, more preferably 95% by weight or more, further preferably 97.5% by weight or more, particularly preferably 99% by weight or more, and most preferably 100% by weight (whole amount). Therefore, it is most preferable that the cell culture scaffold material is the synthetic resin X.
  • the content of the synthetic resin X is the above lower limit or more, the effect of the present invention can be even more effectively exhibited.
  • the cell culture scaffold material may contain components other than the synthetic resin X.
  • Components other than the synthetic resin X include polyolefin resins, polyether resins, polyvinyl alcohol resins, polyesters, epoxy resins, polyamide resins, polyimide resins, polyurethane resins, polycarbonate resins, polysaccharides, celluloses, polypeptides, synthetic peptides, and the like.
  • the content of the component in 100% by weight of the cell culture scaffold material is preferably 10% by weight or less, more preferably 5% by weight or less, further preferably 2.5% by weight or less, particularly preferably 1% by weight or less, and most preferably 0% by weight (not contained). Therefore, it is most preferable that the cell culture scaffold material does not contain any component other than the synthetic resin X.
  • the cell culture scaffold material does not substantially contain animal-derived raw materials. Since it does not contain animal-derived raw materials, it is possible to provide a cell culture scaffold material that is highly safe and has little variation in quality during production.
  • the phrase "does not substantially contain animal-derived raw materials” means that the animal-derived raw materials in the cell culture scaffold material are 3% by weight or less.
  • the animal-derived raw materials in the cell culture scaffold material are preferably 1% by weight or less, and more preferably 0% by weight. That is, it is more preferable that the cell culture scaffold material does not contain any animal-derived raw materials.
  • the cell culture scaffold material is used for culturing cells.
  • the cell culture scaffold material is used as a scaffold for cells when culturing the cells. Therefore, a resin film formed of the cell culture scaffold material of the present invention is used for culturing cells, and is also used as a scaffold for cells when culturing the cells.
  • Examples of the cells include cells of animals such as human, mouse, rat, pig, cow and monkey.
  • examples of the cells include somatic cells and the like, and examples thereof include stem cells, progenitor cells, mature cells, and the like.
  • the somatic cells may be cancer cells.
  • Examples of the mature cells include nerve cells, cardiomyocytes, retinal cells, hepatocytes, and the like.
  • stem cells examples include mesenchymal stem cells (MSCs), iPS cells, ES cells, Muse cells, embryonic cancer cells, embryonic germ cells, mGS cells, and the like.
  • the resin film of the present invention is formed of a cell culture scaffold material.
  • the resin film is formed by using a cell culture scaffold material.
  • the resin film is preferably a film-like cell culture scaffold material.
  • the resin film is preferably a film-like material made of cell culture scaffold material.
  • particles, fibers, a porous body, or a film containing the cell culture scaffold material are also provided.
  • the form of the cell culture scaffold material is not particularly limited, and may be particles, fibers, a porous body, or a film.
  • the particles, fibers, porous body, or film may contain components other than the cell culture scaffold material.
  • the film containing the cell culture scaffold material is preferably used for plane culture (two-dimensional culture) of cells.
  • particles, fibers, or a porous body containing the cell culture scaffold material are preferably used for three-dimensional culture of cells.
  • the present invention also relates to a cell culture carrier in which the resin film is arranged on a surface of the carrier.
  • the cell culture carrier of the present invention can be obtained, for example, by arranging the resin film on the surface of the carrier by coating or the like.
  • the form of the carrier may be particles, fibers, a porous body, or a film. That is, the cell culture carrier of the present invention may be in the form of particles, fibers, a porous body, or a film.
  • the cell culture carrier of the present invention may contain components other than the carrier and the resin film.
  • the present invention also relates to a cell culture vessel including the resin film in at least a part of the cell culture area.
  • Fig. 1 is a cross-sectional view schematically showing a cell culture vessel according to an embodiment of the present invention.
  • a cell culture vessel 1 includes a vessel body 2 and a resin film 3 formed of a cell culture scaffold material.
  • the resin film 3 is arranged on a surface 2a of the vessel body 2.
  • the resin film 3 is arranged on a bottom surface of the vessel body 2.
  • Cells can be cultured in plane by adding a liquid medium to the cell culture vessel 1 and seeding cells on a surface of the resin film 3.
  • the vessel body may include a first vessel body, and a second vessel body such as a cover glass on the bottom surface of the first vessel body.
  • the first vessel body and the second vessel body may be separable.
  • a resin film formed of the cell culture scaffold material may be arranged on the surface of the second vessel body.
  • a conventionally known vessel body can be used.
  • Shape and size of the vessel body are not particularly limited.
  • Examples of the vessel body include a cell culture plate provided with one or a plurality of wells (holes), a cell culture flask, and the like.
  • the number of wells in the plate is not particularly limited.
  • the number of wells is not particularly limited, and examples thereof include 2, 4, 6, 12, 24, 48, 96, 384, and the like.
  • Shape of the well is not particularly limited, and examples thereof include a perfect circle, an ellipse, a triangle, a square, a rectangle, a pentagon, and the like.
  • Shape of the bottom surface of the well is not particularly limited, and examples thereof include a flat bottom, a round bottom, unevenness, and the like.
  • Material of the vessel body is not particularly limited, and examples thereof include resins, metals, and inorganic materials.
  • the resin include polystyrene, polyethylene, polypropylene, polycarbonate, polyester, polyisoprene, cycloolefin polymer, polyimide, polyamide, polyamideimide, (meth)acrylic resin, epoxy resin, silicone, and the like.
  • the metal include stainless steel, copper, iron, nickel, aluminum, titanium, gold, silver, platinum, and the like.
  • the inorganic material include silicon oxide (glass), aluminum oxide, titanium oxide, zirconium oxide, iron oxide, silicon nitride, and the like.
  • the following synthetic resins were synthesized as raw materials for cell culture scaffold material.
  • a reactor equipped with a stirrer was charged with 2700 mL of ion-exchanged water, 300 parts by weight of polyvinyl alcohol with an average degree of polymerization of 1700 and a degree of saponification of 98 mol%, followed by dissolution by heating with stirring to obtain a solution.
  • 35% by weight hydrochloric acid as a catalyst was added such that the concentration of hydrochloric acid became 0.2% by weight.
  • temperature was adjusted to 15°C, and 22 parts by weight of n-butyraldehyde was added thereto with stirring. Then, 148 parts by weight of n-butyraldehyde was added to precipitate a white particulate polyvinyl butyral resin.
  • the obtained synthetic resin has a degree of acetalization (degree of butyralization) of 69 mol%, an amount of hydroxyl groups of 27.5 mol%, a degree of acetylation of 2.0 mol%, a content of vinylpyrrolidone group of 0.3 mol%, and a content of n-lauryl methacrylate of 1.2 mol%.
  • Synthetic resins were obtained in the same manner as in Example 1 except that the weight ratios of the polyvinyl butyral resin, N-vinylpyrrolidone, and n-lauryl methacrylate were changed.
  • Table 1 Table 2 and Table 4 show degrees of acetalization (degrees of butyralization), amounts of hydroxyl groups, degrees of acetylation, contents of vinylpyrrolidone groups, and contents of n-lauryl methacrylate of the synthetic resins obtained in Examples 2 to 11 and Comparative Example 1.
  • a reactor equipped with a stirrer was charged with 2700 mL of ion-exchanged water, 300 parts by weight of polyvinyl alcohol with an average degree of polymerization of 1700 and a degree of saponification of 99 mol%, followed by dissolution by heating with stirring to obtain a solution.
  • 35% by weight hydrochloric acid as a catalyst was added such that the concentration of hydrochloric acid became 0.2% by weight.
  • temperature was adjusted to 15°C, and 22 parts by weight of n-butyraldehyde was added thereto with stirring.
  • n-butyraldehyde was added thereto to precipitate a white particulate polyvinyl acetal resin (polyvinyl butyral resin).
  • 35% by weight hydrochloric acid was added such that the concentration of hydrochloric acid became 1.8% by weight, and then the mixture was heated to 50°C and kept at 50°C for 2 hours.
  • polyvinyl butyral resin an average degree of polymerization of 1700, a degree of acetalization (degree of butyralization) of 70 mol%, an amount of hydroxyl groups of 27 mol%, and a degree of acetylation of 3 mol%).
  • a linear peptide having an amino acid sequence of Gly-Arg-Gly-Asp-Ser (five amino acid residues, described as GRGDS in the table) was prepared. Ten parts by weight of this peptide and 1 part by weight of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (condensing agent) were added to phosphate buffered saline containing neither calcium nor magnesium so that the final concentration of the peptide is 1 mM to prepare a peptidecontaining solution.
  • this peptidecontaining liquid was added to a spin-coated resin film (polyvinyl acetal resin with a linker formed) and reacted to dehydrate and condense a carboxyl group of the linker and an amino group of Gly of the peptide.
  • a peptide-conjugated polyvinyl acetal resin having a polyvinyl acetal resin portion, a linker portion and a peptide portion was prepared.
  • the obtained peptide-conjugated polyvinyl acetal resin had a degree of acetalization (degree of butyralization) of 69 mol%, an amount of hydroxyl groups of 27 mol%, a degree of acetylation of 3 mol%, a content of carboxyl groups of 0.1 mol%, and a content of peptide portion of 1.0 mol%.
  • a peptide-conjugated polyvinyl acetal resin was prepared in the same manner as in Example 12 except that 85 parts by weight of the polyvinyl acetal resin and 15 parts by weight of acrylic acid (linker) were used in the introduction of linker, and the amount of the peptide added was changed to 15 parts by weight in the formation of peptide.
  • a peptide-conjugated polyvinyl acetal resin was prepared in the same manner as in Example 12 except that 70 parts by weight of the polyvinyl acetal resin and 30 parts by weight of acrylic acid (linker) were used in the introduction of linker, and the amount of the peptide added was changed to 30 parts by weight in the formation of peptide.
  • a peptide-conjugated polyvinyl acetal resin was prepared in the same manner as in Example 12 except that 67 parts by weight of the polyvinyl acetal resin and 33 parts by weight of acrylic acid (linker) were used in the introduction of linker, and the amount of the peptide added was changed to 33 parts by weight in the formation of peptide.
  • a peptide-conjugated polyvinyl acetal resin was prepared in the same manner as in Example 12 except that 63 parts by weight of the polyvinyl acetal resin and 37 parts by weight of acrylic acid (linker) were used in the introduction of linker, and the amount of the peptide added was changed to 37 parts by weight in the formation of peptide.
  • a peptide-conjugated polyvinyl acetal resin was prepared in the same manner as in Example 12 except that 30 parts by weight of polyvinyl acetal resin and 70 parts by weight of acrylic acid (linker) were used in the introduction of linker, and the amount of the peptide added was changed to 70 parts by weight in the formation of peptide.
  • a polystyrene resin was used as it was.
  • a reactor equipped with a stirrer was charged with 2700 mL of ion-exchanged water, 300 parts by weight of polyvinyl alcohol with an average degree of polymerization of 1700 and a degree of saponification of 98 mol%, followed by dissolution by heating with stirring to obtain a solution.
  • 35% by weight hydrochloric acid as a catalyst was added such that the concentration of hydrochloric acid became 0.2% by weight.
  • temperature was adjusted to 15°C, and 22 parts by weight of n-butyraldehyde was added thereto with stirring. Then, 148 parts by weight of n-butyraldehyde was added to precipitate a white particulate polyvinyl butyral resin.
  • Vitronectin A laminate of Vitronectin and the cover glass was arranged on a ⁇ 22 mm polystyrene dish to obtain a cell culture vessel. Since Vitronectin is denatured when dried and its adhesive performance is significantly reduced, the cell culture vessel was immersed in a PBS solution immediately after being prepared.
  • Storage elastic moduli at 25°C and 100°C of each scaffold material were measured by a dynamic viscoelasticity measuring device (manufactured by IT Keisoku Seigyo Co., Ltd., DVA-200) under tensile conditions at a frequency of 10 Hz and a temperature range of -150°C to 150°C at a heating rate of 5°C/min.
  • the storage elastic moduli at 25°C and 100°C were obtained from a graph of the obtained tensile storage elastic modulus, and 25°C storage elastic modulus/100°C storage elastic modulus was calculated.
  • the measurement was performed using a measurement sample with a length of 50 mm, a width of 5 to 20 mm, and a thickness of 0.1 to 1.0 mm, under conditions of 10 Hz, a strain of 0.1%, a temperature of -150°C to 150°C, and a heating rate of 5°C/min.
  • a resin solution was obtained by dissolving 1 g of the obtained synthetic resin in 19 g of butanol.
  • the obtained resin solution (150 ⁇ L) was discharged onto a ⁇ 22 mm cover glass (manufactured by Matsunami Glass Ind., Ltd., using 22 round No. 1 after removing dust with an air duster), and rotated at 2000 rpm for 20 seconds using a spin coater to obtain a smooth resin film.
  • the obtained resin film together with the cover 26 glass was arranged on a ⁇ 22 mm polystyrene dish to obtain a cell culture vessel.
  • a laminate of the obtained peptide-conjugated polyvinyl acetal resin and the cover glass was arranged on a ⁇ 22 mm polystyrene dish to obtain a cell culture vessel.
  • the surface free energy of the resin film obtained in the section of preparation of cell culture vessel was measured using a contact angle meter (DMo-701 manufactured by Kyowa Interface Science Co., Ltd.).
  • a contact angle of pure water was obtained by dropping 1 ⁇ L of pure water onto the resin film and photographing a droplet image 30 seconds later.
  • a contact angle of diiodomethane was obtained by dropping 1 ⁇ L of diiodomethane onto the resin film, and photographing a droplet image 30 seconds later.
  • dispersion term component ⁇ d (dSFE) of surface free energy and dipole component ⁇ p (pSFE) as a polar term component of surface free energy were calculated using the Kaelble-Uy theoretical formula.
  • the resin film obtained in the section of preparation of cell culture vessel was observed with an atomic force microscope (AFM, manufactured by Bruker, product number "Dimension XR").
  • AFM atomic force microscope
  • SCAN ASYST AIR was used as a cantilever.
  • Fig. 2 in the resin film of Example 3, a sea-island structure was observed in which the polyvinyl butyral resin portion as the first phase formed a sea part, and a resin portion having the (meth)acrylic acid ester and the vinyl compound (copolymer portion of N-vinylpyrrolidone and n-laurylmethacrylate) as the second phase formed island parts.
  • Examples 1 to 2, 4 to 11 a sea-island structure was observed in which the polyvinyl butyral resin portion as the first phase formed a sea part, and a resin portion having the (meth)acrylic acid ester and the vinyl compound (copolymer portion of N-vinylpyrrolidone and n-laurylmethacrylate) as the second phase formed island parts. Also in Examples 12 to 17, a sea-island structure was observed in which the polyvinyl butyral resin portion as the first phase formed a sea part and the peptide portion as the second phase formed island parts. On the other hand, in Comparative Examples 1 to 4, no phase-separated structure was observed.
  • the ratio of the surface area of the second phase to the entire surface surface area fraction of the phase-separated structure
  • a ratio of the peripheral length to the area of the second phase peripheral length/area
  • the number of the second phases as island parts number of islands
  • the average diameter of the island parts average island size
  • Phosphate buffered saline (1 mL) was added to the obtained cell culture vessel, and the mixture was allowed to stand in an incubator at 37°C for 1 hour, then the phosphate buffered saline in the culture vessel was removed. Colonies of h-iPS cells 253G1 in a confluent state were added to a 35 mm dish, followed by addition of 1 mL of a 0.5 mM ethylenediamine/phosphate buffer solution, and the mixture was allowed to stand at room temperature for 2 minutes.
  • a cell mass (0.5 ⁇ 10 5 cells) crushed to 50 to 200 ⁇ m by pipetting with 1 mL of TeSRE8 medium was seeded in a culture vessel.
  • TeSRE8 medium
  • the cells were cultured in an incubator at 37°C and a CO 2 concentration of 5%.
  • the medium (1 mL) was removed every 24 hours, and 1 mL of fresh TeSR E8 was added to replace the medium.
  • a colonized cell mass after 5 days was exfoliated with 1.0 mL of TryPLE Express exfoliating solution, and the number of cells was counted using a cell counter (Nucleocounter NC-3000, manufactured by Chemometec).
  • a cell proliferation rate relative to Reference Example A was determined using the following formula.
  • the cell proliferation rate was evaluated according to the following criteria.
  • Example 1 Example 2
  • Example 3 Example 4
  • Example 5 Synthetic resin Polyvinyl acetal resin Degree of acetalization (mol%) 69.0 63.0 49.0 39.9 35.0 28.0 Amount of hydroxyl groups (mol%) 27.5 25.2 19.6 16.0 14.0 11.2 Amount of Acetyl groups (mol%) 2.1 1.8 1.4 1.1 1.0 0.8 Vinyl compound A (Acrylic/Vinyl) N-Vinylpyrrolidone (mol%) 0.3 2.0 5.0 7.0 8.3 10.0 n-Lauryl methacrylate (mol%) 1.2 8.0 25.0 36.0 41.7 50.0 Physical properties Storage elastic modulus 25°C Storage elastic modulus (Pa) 2.2 ⁇ 10 9 1.8 ⁇ 10 9 1.9 ⁇ 10 9 1.3 ⁇ 10 9 9.6 ⁇ 10 8 9.0 ⁇ 10 8 100°C Storage elastic modulus (Pa) 2.7 ⁇ 10 6 2.7 ⁇ 10 6 3.0 ⁇ 10 6 4.7 ⁇ 10 6 4.6

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WO2022168871A1 (fr) * 2021-02-03 2022-08-11 積水化学工業株式会社 Microsupport de culture cellulaire et procédé de culture cellulaire
JP2023064377A (ja) * 2021-10-26 2023-05-11 住友化学株式会社 細胞培養基材
WO2023127779A1 (fr) * 2021-12-27 2023-07-06 積水化学工業株式会社 Solution de revêtement pour former un échafaudage cellulaire et son procédé de production
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Family Cites Families (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2400063A1 (fr) * 1977-08-08 1979-03-09 Pasteur Institut Procede d'obtention de supports pour cultures cellulaires et supports obtenus
JP2817934B2 (ja) * 1989-02-28 1998-10-30 旭光学工業株式会社 細胞分離材及び分離器
EP0529751A1 (fr) * 1991-08-09 1993-03-03 W.R. Grace & Co.-Conn. Support et composition d'essai pour culture cellulaire
JP2001089574A (ja) * 1999-07-15 2001-04-03 Kuraray Co Ltd ポリビニルアルコール系含水ゲル、その製造方法及び排水処理装置
JP2003174869A (ja) * 2001-09-20 2003-06-24 Sanyo Chem Ind Ltd 多機能性臓器細胞の接着培養用基材
US7041506B2 (en) * 2001-11-19 2006-05-09 Becton Dickinson And Company Peptides promoting cell adherence, growth and secretion
JP2004033136A (ja) * 2002-07-05 2004-02-05 Fuji Photo Film Co Ltd 細胞培養担体
JP4219361B2 (ja) * 2003-04-25 2009-02-04 積水化学工業株式会社 血液凝固促進剤及び採血管
US7074615B2 (en) * 2003-08-15 2006-07-11 Becton, Dickinson And Company Peptides for enhanced cell attachment and cell growth
JP2006042794A (ja) * 2004-06-28 2006-02-16 Sanyo Chem Ind Ltd 細胞培養用樹脂ビーズ
JP2006314285A (ja) 2005-05-13 2006-11-24 Kuraray Co Ltd 細胞培養用担体及び該細胞培養用担体を用いた細胞培養方法
JP5539649B2 (ja) * 2005-12-13 2014-07-02 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ 細胞移植のための足場
JP4944449B2 (ja) * 2006-01-18 2012-05-30 日東電工株式会社 多孔質構造体の製造方法および多孔質構造体並びに多孔質構造体からなる細胞培養用足場基材
AU2007211018B2 (en) 2006-01-27 2013-09-05 The Regents Of The University Of California Biomimetic scaffolds
JP5441693B2 (ja) * 2007-07-05 2014-03-12 日産化学工業株式会社 新規脂質ペプチド並びにヒドロゲル
JP5000439B2 (ja) * 2007-09-19 2012-08-15 三洋化成工業株式会社 細胞培養用担体
WO2010148346A2 (fr) * 2009-06-19 2010-12-23 The Regents Of The University Of California Matrice d'adhésion cellulaire tridimensionnelle
GB2473814B (en) * 2009-09-16 2014-06-11 Spheritech Ltd Hollow particulate support
JP5846584B2 (ja) * 2010-08-31 2016-01-20 学校法人東京女子医科大学 細胞培養用温度応答性基材及びその製造方法
US10400210B2 (en) * 2012-02-02 2019-09-03 Corning Incorporated Synthetic attachment medium for cell culture
CN202643702U (zh) * 2012-06-07 2013-01-02 江阴瑞康健生物医学科技有限公司 双相多孔三维细胞培养支架
CN102719391A (zh) * 2012-06-07 2012-10-10 江阴瑞康健生物医学科技有限公司 双相多孔三维细胞培养支架
JP2014117268A (ja) * 2012-12-19 2014-06-30 Kyoto Institute Of Technology 修飾ポリペプチドを用いた培養面のコーティング
JP6143163B2 (ja) * 2013-03-22 2017-06-07 国立大学法人三重大学 弾性組織様構造体の製造方法
TWI601817B (zh) * 2013-10-02 2017-10-11 國立中央大學 細胞培養製品及其製造方法
JP5831666B1 (ja) * 2014-12-12 2015-12-09 東洋インキＳｃホールディングス株式会社 細胞培養用部材
JP2016202172A (ja) * 2015-04-16 2016-12-08 国立大学法人京都大学 疑似膵島の製造方法
JP2017023008A (ja) 2015-07-16 2017-02-02 国立大学法人 東京医科歯科大学 ポリロタキサンブロック共重合体表面を有する培養器を用いた幹細胞の培養方法
TW201734124A (zh) * 2015-12-18 2017-10-01 Sekisui Chemical Co Ltd 無機質燒結體製造用黏合劑
JP2018064542A (ja) 2016-10-21 2018-04-26 味の素株式会社 フィブロイン様タンパク質改変体および細胞培養方法
JP7039308B2 (ja) * 2017-02-08 2022-03-22 三洋化成工業株式会社 細胞培養用担体
JPWO2018181758A1 (ja) * 2017-03-31 2020-02-06 積水化学工業株式会社 合わせガラス用中間膜及び合わせガラス
WO2019131982A1 (fr) * 2017-12-27 2019-07-04 積水化学工業株式会社 Matériau de charpente pour cultures de cellules souches et procédé de culture de cellules souches l'utilisant
JP2019118345A (ja) * 2017-12-27 2019-07-22 積水化学工業株式会社 幹細胞培養用足場材料及びそれを用いた幹細胞培養方法
US20220177825A1 (en) * 2019-03-29 2022-06-09 Sekisui Chemical Co., Ltd. Scaffold material for cell culture and cell culture container
EP3950918A4 (fr) * 2019-03-29 2022-12-28 Sekisui Chemical Co., Ltd. Matériau de construction de culture cellulaire, récipient de culture cellulaire, fibre de culture cellulaire et procédé de culture cellulaire
EP3950925A4 (fr) * 2019-03-29 2022-12-28 Sekisui Chemical Co., Ltd. Matériau d'échafaudage de culture cellulaire, récipient de culture cellulaire, support de culture cellulaire, fibre de culture cellulaire et procédé de culture cellulaire

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Ipc: C07K 5/11 20060101ALI20231017BHEP

Ipc: C12M 1/00 20060101ALI20231017BHEP

Ipc: C12N 5/00 20060101AFI20231017BHEP